【摘要】 目的探讨羟基喜树碱(HCPT)对人骨髓增生异常综合征(MDS)细胞系MUTZ-1细胞凋亡中生存素(survivin)基因及蛋白质表达的影响。方法 MUTZ-1细胞与0、1、2、4μg/mL HCPT共育6 h,用流式细胞术(FCM)检测其细胞凋亡率,RT-PCR检测survivin mRNA的表达水平,免疫细胞化学及western blot检测survivin蛋白质的表达水平。结果 MUTZ-1细胞与0、1、2、4μg/mL HCPT共育后,细胞凋亡率升高,分别为(8.58±0.18)%、(12.10±1.04)%、(14.49±0.87)%和(27.34±1.38)%(F=158.7,P<0.05);survivin mRNA表达水平下调,分别为0.87±0.08、0.76±0.03、0.67±0.03和0.45±0.04(F=97.65,P<0.05);细胞凋亡率与survivin mRNA表达水平呈负相关(r=-0.85,P<0.01);survivin蛋白质积分值降低,分别为2.36±0.19、1.50±0.11、1.49±0.18和1.06±0.04(F=14.57,P<0.05);survivin蛋白质表达减少,分别为0.80±0.09、0.34±0.04、0.25±0.02和0.05±0.01(F=37.42,P<0.05)。结论 HCPT可能通过下调survivin基因转录及蛋白质表达,诱导MUTZ-1细胞凋亡。更多还原

【Abstract】 Objective To investigate the effect of hydroxycamptothecine( HCPT) on the expressions of survivin gene and protein during the apoptosis of MUTZ-1 cells,a human myelodysplastic syndrome( MDS) cell line. Methods After MUTZ-1 cells were incubated with HCPT( 0,1,2,4 μg / mL) for 6 h,their apoptotic rates were analyzed by flow cytometry( FCM). Meanwhile,the level of survivin mRNA in MUTZ cells was determined by reverse transcription-polymerase chain reaction( RT-PCR),and the level of survivin protein by immunocytochemistry and western blot. Results After MUTZ-1 cells were incubated with 0,1,2 and 4 μg / mL of HCPT for 6 h,their apoptotic rates were( 8. 58 ± 0. 18) %,( 12. 10 ± 1. 04) %,( 14. 49 ± 0. 87) % and( 27. 34 ± 1. 38) %,respectively, the levels of survivin / β-actin mRNAs 0. 87 ± 0. 08,0. 76 ± 0. 03,0. 67 ± 0. 03 and 0. 45 ± 0. 04,the integral values 2. 36 ± 0. 19, 1. 50 ± 0. 11,1. 49 ± 0. 18 and 1. 06 ± 0. 04,and the levels of survivin / β-actin proteins 0. 80 ± 0. 09,0. 34 ± 0. 04,0. 25 ± 0. 02 and 0. 05 ± 0. 01,respectively. Compared with the control( 0 μg / mL HCPT),the apoptotic rates of the other three groups increased( F = 158. 7,P < 0. 05),and the levels of survivin / β-actin mRNAs,the integral values and the levels of survivin / β-actin proteins decreased( F =97. 65,14. 57 and 37. 42,respectively,P <0. 05). There was a negative correlation between the apoptotic rate and the level of survivin mRNA expression( r =- 0. 85,P < 0. 01). Conclusion HCPT induced the apoptosis of MUTZ-1 cells probably by downregulating the expressions of survivin mRNA and protein.更多还原